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利用任意引物聚合酶链反应对牙龈卟啉单胞菌进行克隆分析。

Clonal analysis of Porphyromonas gingivalis by the arbitrarily primed polymerase chain reaction.

作者信息

Chen C, Slots J

机构信息

Department of Periodontology, USC School of Dentistry, Los Angeles, California.

出版信息

Oral Microbiol Immunol. 1994 Apr;9(2):99-103. doi: 10.1111/j.1399-302x.1994.tb00042.x.

Abstract

Genetic analysis of Porphyromonas gingivalis strains may distinguish between virulent and nonvirulent strains and also may be used to trace individual strains in epidemiological studies. The present study examined the utility of the arbitrarily primed polymerase chain reaction for genotypic fingerprinting of P. gingivalis. DNA was extracted according to conventional methods. Ten-base oligonucleotide primers with arbitrary sequences were used with the polymerase chain reaction to amplify P. gingivalis genomic DNA. The amplification products were analyzed by agarose gel electrophoresis. The primer GACCGCTTGT grouped 73 P. gingivalis strains into 23 genotypes, including 16 genotypes containing a single strain each. The primer AGGGGTCTTG identified 45 different genotypes, 33 of which contained a single strain. P. gingivalis strains ATCC 33277T and 381 belonged to the same genotype. Likewise, strains W50 and W83 were of the same genetic clone. The present study indicates that the arbitrarily primed polymerase chain reaction represents a valuable and easy method for clonal analysis of P. gingivalis.

摘要

牙龈卟啉单胞菌菌株的基因分析可以区分有毒菌株和无毒菌株,也可用于在流行病学研究中追踪单个菌株。本研究检测了任意引物聚合酶链反应用于牙龈卟啉单胞菌基因分型的实用性。按照常规方法提取DNA。使用具有任意序列的10碱基寡核苷酸引物,通过聚合酶链反应扩增牙龈卟啉单胞菌基因组DNA。扩增产物通过琼脂糖凝胶电泳进行分析。引物GACCGCTTGT将73株牙龈卟啉单胞菌分为23种基因型,其中16种基因型各包含一个菌株。引物AGGGGTCTTG鉴定出45种不同的基因型,其中33种包含一个菌株。牙龈卟啉单胞菌菌株ATCC 33277T和381属于同一基因型。同样,菌株W50和W83属于同一基因克隆。本研究表明,任意引物聚合酶链反应是一种用于牙龈卟啉单胞菌克隆分析的有价值且简便的方法。

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